Biotin Conjugated Human GDF-15 Recombinant Rabbit Monoclonal Antibody [PSH16-01] - Detector

Sandwich ELISA analysis of Human GDF15 matched pair antibodies

Capture: HA723805, Human GDF-15 Rabbit mAb [PSH16-00]
Detector: HA723806, Human GDF-15 Rabbit mAb [PSH16-01]

Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA723805) diluted in carbonate/bicarbonate buffer, at a concentration of 2ug/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1% BSA blocking buffer, and incubated with serial diluted Recombinant Human GDF-15 protein starting from 3000 pg/ml to 0 pg/ml and detect antibody (HA723806, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

Interpolated concentrations of native GDF-15 in human samples.

Capture: HA723805, Human GDF-15 Rabbit mAb [PSH16-00]
Detector: HA723806, Human GDF-15 Rabbit mAb [PSH16-01]

Interpolated concentration of native GDF-15 was measured in duplicate at different sample concentrations. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean GDF-15 concentration was determined to be 17,760 pg/mL in HepG2 and 1,208 pg/ml in JAR cell culture supernatant . There was no detectable signal in U-937 cell culture supernatant.

Interpolated concentrations of spiked GDF-15 in cell culture media samples.

Capture: HA723805, Human GDF-15 Rabbit mAb [PSH16-00]
Detector: HA723806, Human GDF-15 Rabbit mAb [PSH16-01]

The concentrations of GDF-15 were measured in duplicates, interpolated from the GDF-15 standard curves and corrected for sample dilution. Diluted samples are as follows: 50% cell culture media with FBS.The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).